The present invention relates to novel compounds, to methods for their preparation, to compositions comprising the compounds, to the use of these compounds as medicaments and their use in therapy, where such compounds of Formula 1 are pharmacologically useful inhibitors of Protein Tyrosine Phosphatases (PTPases) such as PTP1B, CD45, SHP-1, SHP-2, PTPxcex1, LAR and HePTP or the like, 
wherein n, m, X, Y, R1, R2, R3, R4, R5 and R6 are defined more fully below. It has been found that PTPases plays a major role in the intracellular modulation and regulation of fundamental cellular signaling mechanisms involved in metabolism, growth, proliferation and differentiation (Hunter, Phil. Trans. R. Soc. Lond. B 353: 583-605 (1998); Chan et al., Annu. Rev. Immunol. 12: 555-592 (1994); Zhang, Curr. Top. Cell. Reg. 35: 21-68 (1997); Matozaki and Kasuga, Cell. Signal. 8: 113-19 (1996); Flint et al., The EMBO J. 12:1937-46 (1993); Fischer et al, Science 253:401-6 (1991)). Overexpression or altered activity of tyrosine phosphatases can also contribute to the symptoms and progression of various diseases (Wiener, et al., J. Natl. cancer Inst. 86:372-8 (1994); Hunter and Cooper, Ann. Rev. Biochem, 54:897-930 (1985)). Furthermore, there is increasing evidence which suggests that inhibition of these PTPases may help treat certain types of diseases such as diabetes type I and II , autoimmune disease, acute and chronic inflammation, osteoporosis and various forms of cancer.
Protein phosphorylation is now well recognized as an important mechanism utilized by cells to transduce and regulate signals during different stages of cellular function (Hunter, Phil. Trans. R. Soc. Lond. B 353: 583-605 (1998); Chan et al., Annu. Rev. Immunol. 12: 555-592 (1994); Zhang, Curr. Top. Cell. Reg. 35: 21-68 (1997); Matozaki and Kasuga, Cell. Signal. 8: 113-19 (1996); Fischer et al, Science 253:401-6 (1991); Flint et al., EMBO J. 12:1937-46 (1993)). There are at least two major classes of phosphatases: (1) those that dephosphorylate proteins (or peptides) that contain a phosphate group(s) on a serine or threonine moiety (termed Ser/Thr phosphatases) and (2) those that remove a phosphate group(s) from the amino acid tyrosine (termed protein tyrosine phosphatases or PTPases or PTPs).
The PTPases are a family of enzymes that can be classified into two groups: a) intracellular or nontransmembrane PTPases and b) receptor-type or transmembrane PTPases.
Intracellular PTPases
Most known intracellular type PTPases contain a single conserved catalytic phosphatase domain consisting of 220-240 amino acid residues. The regions outside the PTPase domains are believed to play important roles in localizing the intracellular PTPases subcellularly (Mauro, L. J. and Dixon, J. E. TIBS 19: 151-155 (1994)). The first intracellular PTPase to be purified and characterized was PTP1B, which was isolated from human placenta (Tonks et al., J. Biol. Chem. 263: 6722-6730 (1988)). Shortly after, PTP1B was cloned (Charbonneau et al., Proc. Natl. Acad. Sci. USA 86: 5252-5256 (1989); Chernoff et al., Proc. Natl. Acad. Sci. USA 87: 2735-2789 (1989)). Other examples of intracellular PTPases include (1) T-cell PTPase/TC-PTP (Cool et al. Proc. Natl. Acad. Sci. USA 86: 5257-5261 (1989)), (2) rat brain PTPase (Guan et al., Proc. Natl. Acad. Sci. USA 87:1501-1502 (1990)), (3) neuronal phosphatase STEP (Lombroso et al., Proc. Natl. Acad. Sci. USA 88: 7242-7246 (1991)), (4) ezrin-domain containing PTPases: PTPMEG1 (Guet al., Proc. Natl. Acad. Sci. USA 88: 5867-57871 (1991)), PTPH1 (Yang and Tonks, Proc. Natl. Acad. Sci. USA 88: 5949-5953 (1991)), PTPD1 and PTPD2 (Mxc3x8ller et al., Proc. Natl. Acad. Sci. USA 91: 7477-7481 (1994)), FAP-1/BAS (Sato et al., Science 268: 411-415 (1995); Banville et al., J. Biol. Chem. 269: 22320-22327 (1994); Maekawa et al., FEBS Letters 337: 200-206 (1994)), and SH2 domain containing PTPases: PTP1C/SH-PTP1/SHP-1 (Plutzky et al., Proc. Natl. Acad. Sci. USA 89: 1123-1127 (1992); Shen et al., Nature Lond. 352: 736-739 (1991)) and PTP1D/Syp/SH-PTP2/SHP-2 (Vogel et al., Science 259: 1611-1614 (1993); Feng et al., Science 259: 1607-1611 (1993); Bastein et al., Biochem. Biophys. Res. Comm. 196: 124-133 (1993)).
Receptor-type PTPases consist of a) a putative ligand-binding extracellular domain, b) a transmembrane segment, and c) an intracellular catalytic region. The structures and sizes of the putative ligand-binding extracellular domains of receptor-type PTPases are quite divergent. In contrast, the intracellular catalytic regions of receptor-type PTPases are very homologous to each other and to the intracellular PTPases. Most receptor-type PTPases have two tandemly duplicated catalytic PTPase domains.
The first receptor-type PTPases to be identified were (1) CD45/LCA (Ralph, S. J., EMBO J. 6: 1251-1257 (1987)) and (2) LAR (Streuli et al., J. Exp. Med. 168: 1523-1530 (1988)) that were recognized to belong to this class of enzymes based on homology to PTP1B (Charbonneau et al., Proc. Natl. Acad. Sci. USA 86: 5252-5256 (1989)). CD45 is a family of high molecular weight glycoproteins and is one of the most abundant leukocyte cell surface glycoproteins and appears to be exclusively expressed upon cells of the hematopoietic system (Trowbridge and Thomas, Ann. Rev. Immunol. 12: 85-116 (1994)).
The identification of CD45 and LAR as members of the PTPase family was quickly followed by identification and cloning of several different members of the receptor-type PTPase group. Thus, 5 different PTPases, (3) PTPxcex1, (4) PTPxcex2, (5) PTPxcex4, (6) PTPxcex5, and (7) PTPxcex6, were identified in one early study (Krueger et al., EMBO J. 9: 3241-3252 (1990)). Other examples of receptor-type PTPases include (8) PTPxcex3 (Barnea et al., Mol. Cell. Biol. 13: 1497-1506 (1995)) which, like PTPxcex6 (Krueger and Saito, Proc. Natl. Acad. Sci. USA 89: 7417-7421 (1992)) contains a carbonic anhydrase-like domain in the extracellular region, (9) PTPxcexc (Gebbink et al., FEBS Letters 290: 123-130 (1991)), (10) PTPxcexa (Jiang et al., Mol. Cell. Biol. 13: 2942-2951 (1993)). Based on structural differences the receptor-type PTPases may be classified into subtypes (Fischer et al., Science 253: 401-406 (1991)): (I) CD45; (II) LAR, PTPxcex4, (11) PTP"sgr"; (III) PTP, (12) SAP-1 (Matozaki et al., J. Biol. Chem. 269: 2075-2081 (1994)), (13) PTP-U2/GLEPP1 (Seimiya et al., Oncogene 10: 1731-1738 (1995); Thomas et al., J. Biol. Chem. 269: 19953-19962 (1994)), and (14) DEP-1; (IV) PTPxcex1, PTPxcex5. All receptor-type PTPases except Type III contain two PTPase domains. Novel PTPases are continuously identified. In the early days of PTPase research, it was believed that the number of PTPs would match that of protein tyrosine kinases (PTKs) (Hanks and Hunter, FASEB J. 9: 576-596 (1995)). However, although about 90 open reading frames in C. elegans contain the hallmark motif of PTPs, it now seems that the estimate of xe2x80x98classicalxe2x80x99 PTPases must be downsized, perhaps to between 100 and 200 in humans.
PTPases are the biological counterparts to protein tyrosine kinases Therefore, one important function of PTPases is to control, down-regulate, the activity of PTKs. However, a more complex picture of the function of PTPases has emerged. Thus, several studies have shown that some PTPases may actually act as positive mediators of cellular signaling. As an example, the SH2 domain-containing SHP-2 seems to act as a positive mediator in insulin-stimulated Ras activation (Noguchi et al., Mol. Cell. Biol. 14: 6674-6682 (1994)) and of growth factor-induced mitogenic signal transduction (Xiao et al., J. Biol. Chem. 269: 21244-21248 (1994)), whereas the homologous SHP-1 seems to act as a negative regulator of growth factor-stimulated proliferation (Bignon and Siminovitch, Clin.Immunol. Immunopathol. 73: 168-179 (1994)). Another example of PTPases as positive regulators has been provided by studies designed to define the activation of the Src-family of tyrosine kinases. In particular, several lines of evidence indicate that CD45 is positively regulating the activation of hematopoietic cells, possibly through dephosphorylation of the C-terminal tyrosine of Fyn and Lck (Chan et al., Annu. Rev. Immunol. 12: 555-592 (1994)).
PTPases were originally identified and purified from cell and tissue lysates using a variety of artificial substrates and, therefore, their natural function of dephosphorylation was not well known. Since tyrosine phosphorylation by tyrosine kinases is usually associated with cell proliferation, cell transformation and cell differentiation, it was assumed that PTPases were also associated with these events. This association has now been proven to be the case with many PTPases. PTP1 B, a phosphatase whose structure was the first PTPase to be elucidated (Barford et al., Science 263:1397-1404 (1994)) has been shown to be involved in insulin-induced oocyte maturation (Flint et al., The EMBO J. 12:1937-46 (1993)) and it has been suggested that the overexpression of this enzyme may be involved in p185c-erb B2-associated breast and ovarian cancers (Wiener, et al., J. Natl. cancer Inst. 86:372-8 (1994); Weiner et al., Am. J. Obstet. Gynecol. 170:1177-883 (1994)). The association with cancer is recent evidence which suggests that overexpression of PTP1B is statistically correlated with increased levels of p185c-erb B2 in ovarian and breast cancer. The role of PTP1B in the etiology and progression of the disease has not yet been elucidated. Inhibitors of PTP1B may therefore help clarify the role of PTP1B in cancer and in some cases provide therapeutic treatment for certain forms of cancer.
PTPases: The Insulin Receptor Signaling Pathway/diabetes
Insulin is an important regulator of different metabolic processes and plays a key role in the control of blood glucose. Defects related to its synthesis or signaling lead to diabetes mellitus. Binding of insulin to the insulin receptor (IR) causes rapid (auto)phosphorylation of several tyrosine residues in the intracellular part of the xcex2-subunit. Three closely positioned tyrosine residues (the tyrosine-1150 domain) must all be phosphorylated to obtain full activity of the insulin receptor tyrosine kinase (IRTK) which transmits the signal further downstream by tyrosine phosphorylation of other cellular substrates, including insulin receptor substrate-1 (IRS-1) (Wilden et al., J. Biol. Chem. 267: 16660-16668 (1992); Myers and White, Diabetes 42: 643-650 (1993); Lee and Pilch, Am. J. Physiol. 266: C319-C334 (1994); White et al., J. Biol. Chem. 263: 2969-2980 (1988)). The structural basis for the function of the tyrosine-triplet has been provided by X-ray crystallographic studies of IRTK that showed tyrosine-1150 to be autoinhibitory in its unphosphorylated state (Hubbard et al., Nature 372: 746-754 (1994)) and of the activated IRTK (Hubbard, EMBO J. 16:5572-5581 (1997)).
Several studies clearly indicate that the activity of the auto-phosphorylated IRTK can be reversed by dephosphorylation in vitro (reviewed in Goldstein, Receptor 3: 1-15 (1993); Mooney and Anderson, J. Biol. Chem. 264: 6850-6857 (1989)), with the tri-phosphorylated tyrosine-1150 domain being the most sensitive target for protein-tyrosine phosphatases (PTPases) as compared to the di- and mono-phosphorylated forms (King et al., Biochem. J. 275: 413-418 (1991)). This tyrosine-triplet functions as a control switch of IRTK activity and the IRTK appears to be tightly regulated by PTP-mediated dephosphorylation in vivo (Khan et al., J. Biol. Chem. 264: 12931-12940 (1989); Faure et al., J. Biol. Chem. 267: 11215-11221 (1992); Rothenberg et al., J. Biol. Chem. 266: 8302-8311 (1991)). The intimate coupling of PTPases to the insulin signaling pathway is further evidenced by the finding that insulin differentially regulates PTPase activity in rat hepatoma cells (Meyerovitch et al., Biochemistry 31: 10338-10344 (1992)) and in livers from alloxan diabetic rats (Boylan et al., J. Clin. Invest. 90: 174-179 (1992)).
Until recently, relatively little was known about the identity of the PTPases involved in IRTK regulation. However, the existence of PTPases with activity towards the insulin receptor can be demonstrated as indicated above. Further, when the strong PTPase-inhibitor pervanadate is added to whole cells an almost full insulin response can be obtained in adipocytes (Fantus et al., Biochemistry 28: 8864-8871 (1989); Eriksson et al., Diabetologia 39: 235-242 (1995)) and skeletal muscle (Leighton et al., Biochem. J. 276: 289-292 (1991)). In addition, other studies show that a new class of peroxovanadium compounds act as potent hypoglycemic compounds in vivo (Posner et al.,supra). Two of these compounds were demonstrated to be more potent inhibitors of dephosphorylation of the insulin receptor than of the EGF-receptor, thus indicating that even such relatively unselective inhibitors may convey some specificity in regulating different signal transduction pathways.
It was recently found by Montreal-based research groups that mice lacking the protein tyrosine phosphatase-1B gene (PTP1B) (Elchebly et al., Science 283: 1544-1548 (1999)) yielded healthy mice that showed increased insulin sensitivity and resistance to diet-induced obesity. Importantly, these results have been confirmed and extended independently by another research team from Boston (Klaman et al., Mol. Cell. Biol. 20: 5479-5489 (2000)). The enhanced insulin sensitivity of the PTPxe2x88x92/xe2x88x92 mice was also evident in glucose and insulin tolerance tests. The PTP-1B knock-out mouse showed many characteristics which would be highly desirable to have for an anti-diabetes treatment. Most importantly, the knock-out mice grew normally and were fertile and have exhibited no increased incidence of cancer, as obviously there could have been concerns when one considers the mitogenic properties of insulin. From the diabetes perspective, the first notable features of the knock-out animals were that blood glucose and insulin levels were lowered, and the consequent marked increase in insulin sensitivity in the knock-out animals. Moreover, the insulin-stimulated tyrosine phosphorylation levels of IR and IRS-1 were found to be increased/prolonged in muscle and liverxe2x80x94but not in fat tissue. Thus, the main target tissues for this type of approach would appear to be insulin action in liver and muscle. This is in contrast to the main target tissue for the PPARxcex3 agonist class of insulin sensitizers (the xe2x80x9c-dionesxe2x80x9d), which is adipose tissue (Murphy and Nolan, Exp. Opin. Invest. Drugs 9: 1347-1361 (2000)). Several other xe2x80x9cdiabeticxe2x80x9d parameters were also improved, such as plasma triglycerides being decreased in the knock-out mice. However, perhaps even more remarkably and unexpectedly, the knock-out animals also exhibited a resistance to weight gain when placed on a high-fat diet. This is again in contrast to the action of the PPARxcex3 agonist class of insulin sensitizers, which rather induce weight gain (Murphy and Nolan, supra), and would suggest that inhibition of PTP-1B could be a particularly attractive option for treatment of obese Type II diabetics. This is also supported by the fact that the heterozygous mice from this study showed many of these desirable features. In the Montreal study, there appeared to be no gender differences, whereas in the Boston study in general the male animals had larger responses to PTP-1B being knocked out. In both studies, the reduction in weight gain of the knock-out animals on the high fat diet was found to be due to a decreased fat cell mass, although differences were observed with respect to fat cell number. Leptin levels were also lower in the knock-out mice, presumably as a reflection of the decreased fat mass. Significantly, the Boston group also found that the knock-out animals had an increased energy expenditure of around 20% and an increased respiratory quotient compared to the wild-type; again, heterozygote animals displayed an intermediate level of energy expenditure. Whether this increase in metabolic rate is a reflection of the effects of PTP-1B on insulin-signaling or on other cellular components remains to be established, but the bottom-line message that inhibition of this enzyme may be an effective anti-diabetic and perhaps also anti-obesity therapy is clear.
It should also be noted that in the PTP-1B knock-out mice the basal tyrosine phosphorylation level of the insulin receptor tyrosine kinase does not appear to be increased, which is in contrast to the situation after insulin treatment where there is an increased or prolonged phosphorylation. This might indicate that other PTPs are controlling the basic phosphorylation state of the insulin receptor in the knock-out micexe2x80x94and perhaps in man.
Previous findings are in accordance with the results reported by Elchebly et al. (supra) (recently reviewed in Kennedy, Biomed. Pharmacother. 53: 466-470 (1999)). Thus, it has been found that high glucose concentration induce insulin resistance and increase the expression of PTP1B in rat (fibroblasts expressing the human insulin receptor (Maegawa et al., J. Biol. Chem. 270: 7724-7730 (1995)). In rat L6 cells, insulin and insulin-like growth factor I (IGF-I) were found to induce increased PTPase activity, including increased PTP1B expression (Kenner et al. J. Biol. Chem. 266: 25455-25462 (1993)). In addition, the same group has shown that PTP1B may interact directly with the activated IR (Seely et al. Diabetes 45: 1379-1385 (1996)) and act directly as a negative regulator of insulin and IGF-I-stimulated signaling (Kenner et al. J. Biol. Chem. 271: 19810-19816 (1996)). Osmotic loading of rat KRC-7 hepatoma cells with neutralizing anti-PTP1B antibodies also indicated a role for PTP1B in negative regulating of the insulin signaling pathway (Akmad et al. J. Biol. Chem. 270: 20503-20508 (1995)).
Also other PTPases have been implicated as regulators of the insulin signaling pathway. Thus, it was found that the ubiquitously expressed SH2 domain containing PTPase, PTP1D/SHP-2 (Vogel et al., 1993, supra), associates with and dephosphorylates IRS-1, but apparently not the IR itself (Kuhnxc3xa9 et al., J. Biol. Chem. 268: 11479-11481 (1993); (Kuhnxc3xa9 et al., J. Biol. Chem. 269: 15833-15837 (1994)).
Other studies suggest that receptor-type or membrane-associated PTPases are involved in IRTK regulation (Faure et al., J. Biol. Chem. 267: 11215-11221 (1992), (Hxc3xa4ring et al., Biochemistry 23: 3298-3306 (1984); Sale, Adv. Prot. Phosphatases 6: 159-186 (1991)). Hashimoto et al. have proposed that LAR might play a role in the physiological regulation of insulin receptors in intact cells (Hashimoto et al., J. Biol. Chem. 267: 13811-13814 (1992)). Their conclusion was reached by comparing the rate of dephosphorylation/inactivation of purified IR using recombinant PTP1B as well as the cytoplasmic domains of LAR and PTPxcex1. Antisense inhibition was used to study the effect of LAR on insulin signaling in a rat hepatoma cell line (Kulas et al., J. Biol. Chem. 270: 2435-2438 (1995)). A suppression of LAR protein levels by about 60 percent was paralleled by an approximately 150 percent increase in insulin-induced auto-phosphorylation. However, only a modest 35 percent increase in IRTK activity was observed, whereas the insulin-dependent phosphatidylinositol 3-kinase (PI 3-kinase) activity was significantly increased by 350 percent. Reduced LAR levels did not alter the basal level of IRTK tyrosine phosphorylation or activity. The authors speculate that LAR could specifically dephosphorylate tyrosine residues that are critical for PI 3-kinase activation either on the insulin receptor itself or on a downstream substrate. Conflicting results have been reported for PTP-LAR knock-out mice. Thus, Goldstein and coworkers reported that transgenic mice deficient in PTP-LAR exhibit profound defects in glucose-homeostasis (Ren et al., Diabetes 47: 493-497 (1998)). However, it is difficult to fully assess the contribution of LAR deficiency to the glucose homeostasis in these mice due to the fact that the control mice were of a different genetic background than the knock-out mice. Moreover, normal glucose homeostasis was reported in a different strain of PTP-LAR knock-out mice (Sorensen et al., Diabetologia 40: A143 (1997)).
While previous reports indicate a role of PTPxcex1 in signal transduction through src activation (Zheng et al., Nature 359: 336-339 (1992); den Hertog et al., EMBO J. 12: 3789-3798 (1993)) and interaction with GRB-2 (den Hertog et al., EMBO J. 13: 3020-3032 (1994); Su et al., J. Biol. Chem. 269: 18731-18734 (1994)), Mxc3x8ller, Lammers and coworkers provided results that suggest a function for this phosphatase and its close relative PTP as negative regulators of the insulin receptor signal (Mxc3x8ller et al, 1995 supra;
Lammers, et al., FEBS Lett. 404:37-40 (1997). These studies also indicated that receptor-like PTPases might play a significant role in regulating the IRTK.
Other studies have shown that PTP1B and TC-PTP are likely to be involved in the regulation of several other cellular processes in addition to the described regulatory roles in insulin signaling. Therefore, PTP1B and/or TC-PTP as well as other PTPases showing key structural features with PTP1B and TC-PTP are likely to be important therapeutic targets in a variety of human and animal diseases. The compounds of the present invention are useful for modulating or inhibiting PTP1B and/or TC-PTP and/or other PTPases showing key structural features with said PTPases and for treating diseases in which said modulation or inhibition is indicated. A few examples that are not intended in any way to limit the scope of the invention of substrates that may be regulated by PTP1B will be given below.
Tonks and coworkers have developed an elegant xe2x80x98substrate trappingxe2x80x99 technique that has allowed identification of the epidermal growth factor receptor (EGF-R) as a major substrate of PTP1B in COS cells (Flint et al. Proc. Natl. Acad. Sci. USA 94: 1680-1685 (1997)). In addition, three other as yet unidentified substrates of PTP1B were isolated. As an example of these studies, it has been foundxe2x80x94using the above substrate-trapping techniquexe2x80x94that PTP1B in addition to the EGF-R associates with activated platelet-derived growth factor receptor (PDGF-R), but not with colony-stimulating factor 1 receptor (CSF-1R) (Liu and Chernoff, Biochem. J. 327: 139-145 (1997)).
Early studies have shown that the subcellular localization as well as the enzyme activity of PTP1B may be regulated by agonist-induced calpain-catalyzed cleavage in human platelets (Frangioni et al. EMBO J. 12: 4843-4856 (1993)). Moreover, PTP1B cleavage correlated with the transaction from reversible to irreversible platelet aggregation. Thus, as a non-limiting example compounds of the present invention might be used to prevent or induce irreversible platelet aggregation in individuals in need thereof. It was proposed that the cleavage-induced change in the subcellular localization of PTP1B (from membrane to cytosol) results in different substrate specificity not only in platelet but also in other cell types (Frangioni et al., supra).
The above substrate trapping method has further been used to identify the protein tyrosine kinase p210bcr-abl as a substrate for PTP1B (LaMontagne, Jr. et al. Mol. Cell. Biol. 18: 2965-2975 (1998)). These studies suggest that PTP1B might function as a negative regulator of p210bcr-abl signaling in vivo. In addition, PTP1B was recently found to bind to and dephosphorylate the docking protein p130Cas in rat fibroblasts and thereby suppress transformation by v-crk, v-src, and v-ras, but not by v-raf (Liu et al. Mol. Cell. Biol. 18: 250-259 (1998)).
The transmembrane PTPase CD45, which is believed to be hematopoietic cell-specific, was found to negatively regulate the insulin receptor tyrosine kinase in the human multiple myeloma cell line U266 (Kulas et al., J. Biol. Chem. 271: 755-760 (1996)).
Further, PTPases influences the following hormones or diseases or disease states: somatostatin, the immune system/autoimmunity, cell-cell interactions/cancer, platelet aggregation, osteoporosis, and microorganisms, as disclosed in PCT Publication WO 99/15529.
Somatostatin inhibits several biological functions including cellular proliferation (Lamberts et al., Molec. Endocrinol. 8: 1289-1297 (1994)). While part of the antiproliferative activities of somatostatin are secondary to its inhibition of hormone and growth factor secretion (e.g. growth hormone and epidermal growth factor), other antiproliferative effects of somatostatin are due to a direct effect on the target cells. As an example, somatostatin analogs inhibit the growth of pancreatic cancer presumably via stimulation of a single PTPase, or a subset of PTPases, rather than a general activation of PTPase levels in the cells (Liebow et al., Proc. Natl. Acad. Sci. USA 86: 2003-2007 (1989); Colas et al., Eur. J. Biochem. 207:1017-1024 (1992)).
PTPases: The Immune System/autoimmunity
Several studies suggest that the receptor-type PTPase CD45 plays a critical role not only for initiation of T cell activation, but also for maintaining the T cell receptor-mediated signaling cascade. These studies are reviewed in: (Weiss A., Ann. Rev. Genet. 25: 487-510 (1991); Chan et al., Annu. Rev. Immunol. 12: 555-592 (1994); Trowbridge and Thomas, Annu. Rev. Immunol. 12: 85-116 (1994)).
CD45 is one of the most abundant of the cell surface glycoproteins and is expressed exclusively on hemopoetic cells. In T cells, it has been shown that CD45 is one of the critical components of the signal transduction machinery of lymphocytes. In particular, evidence has suggested that CD45 phosphatase plays a pivotal role in antigen-stimulated proliferation of T lymphocytes after an antigen has bound to the T cell receptor (Trowbridge, Ann. Rev. Immunol, 12: 85-116 (1994)). Several studies suggest that the PTPase activity of CD45 plays a role in the activation of Lck, a lymphocyte-specific member of the Src family protein-tyrosine kinase (Mustelin et al., Proc. Natl. Acad. Sci. USA 86: 6302-6306 (1989); Ostergaard et al., Proc. Natl. Acad. Sci. USA 86: 8959-8963 (1989)). These authors hypothesized that the phosphatase activity of CD45 activates Lck by dephosphorylation of a C-terminal tyrosine residue, which may, in turn, be related to T-cell activation. Thus, it was found that recombinant p56lck specifically associates with recombinant CD45 cytoplasmic domain protein, but not to the cytoplasmic domain of the related PTPxcex1 (Ng et al., J. Biol. Chem. 271: 1295-1300 (1996)). The p56lck-CD45 interaction seems to be mediated via a nonconventional SH2 domain interaction not requiring phosphotyrosine. In immature B cells, another member of the Src family protein-tyrosine kinases, Fyn, seems to be a selective substrate for CD45 compared to Lck and Syk (Katagiri et al., J. Biol. Chem. 270: 27987-27990 (1995)).
Studies using transgenic mice with a mutation for the CD45-exon6 exhibited lacked mature T cells. These mice did not respond to an antigenic challenge with the typical T cell mediated response (Kishihara et al., Cell 74:143-56 (1993)). Inhibitors of CD45 phosphatase would therefore be very effective therapeutic agents in conditions that are associated with autoimmune diseases with rheumatoid arthritis, systemic lupus erythematosus, type I diabetes, and inflammatory bowel disease as non-limiting examples. Another important use of CD45 inhibitors is for immunosuppression in connection with tissue or cell transplantation and other condtions with need for immunosuppressive treatment.
CD45 has also been shown to be essential for the antibody mediated degranulation of mast cells (Berger et al., J. Exp. Med. 180:471-6 (1994)). These studies were also done with mice that were CD45-deficient. In this case, an IgE-mediated degranulation was demonstrated in wild type but not CD45-deficient T cells from mice. These data suggest that CD45 inhibitors could also play a role in the symptomatic or therapeutic treatment of allergic disorders with asthma, allergic rhinitis, food allergy, eczema, urticaria and anaphylaxis as non-limiting examples.
Another PTPase, an inducible lymphoid-specific protein tyrosine phosphatase (HePTP) has also been implicated in the immune response. This phosphatase is expressed in both resting T and B lymphocytes, but not non-hemopoetic cells. Upon stimulation of these cells, mRNA levels from the HePTP gene increase 10-15 fold (Zanke et al., Eur. J. Immunol. 22: 235-239 (1992)). In both T and B cells HePTP may function during sustained stimulation to modulate the immune response through dephosphorylation of specific residues. Its exact role, however remains to be defined.
Likewise, the hematopoietic cell specific SHP-1 seems to act as a negative regulator and play an essential role in immune cell development. In accordance with the above-mentioned important function of CD45, HePTP and SHP-1, selective PTPase inhibitors may be attractive drug candidates both as immunosuppressors and as immunostimulants. Recent studies illustrate the potential of PTPase inhibitors as immunmodulators by demonstrating the capacity of the non-selective vanadium-based PTPase inhibitor, BMLOV, to induce apparent B cell selective apoptosis compared to T cells (Dawson et al., FEBS Lett. 478: 233-236; Schieven et al., J. Biol. Chem. 270: 20824-20831 (1995)).
PTPases: Cell-cell Interactions/cancer
Focal adhesion plaques, an in vitro phenomenon in which specific contact points are formed when fibroblasts grow on appropriate substrates, seem to mimic, at least in part, cells and their natural surroundings. Several focal adhesion proteins are phosphorylated on tyrosine residues when fibroblasts adhere to and spread on extracellular matrix (Gumbiner, Neuron 11: 551-564 (1993)). However, aberrant tyrosine phosphorylation of these proteins can lead to cellular transformation. The intimate association between PTPases and focal adhesions is supported by the finding of several intracellular PTPases with ezrin-like N-terminal domains, e.g. PTPMEG1 (Gu et al., Proc. Natl. Acad. Sci. USA 88: 5867-5871 (1991), PTPH1 (Yang and Tonks, Proc. Natl. Acad. Sci. USA 88: 5949-5953 (1991)) and PTPD1 (Mxc3x8ller et al., Proc. Natl. Acad. Sci. USA 91: 7477-7481 (1994)). The ezrin-like domain shows similarity to several proteins that are believed to act as links between the cell membrane and the cytoskeleton. PTPD1 was found to be phosphorylated by and associated with c-src in vitro and is hypothesized to be involved in the regulation of phosphorylation of focal adhesions (Mxc3x8ller et al., supra).
PTPases may oppose the action of tyrosine kinases, including those responsible for phosphorylation of focal adhesion proteins, and may therefore function as natural inhibitors of transformation. TC-PTP, and especially the truncated form of this enzyme (Cool et al., Proc. Natl. Acad. Sci. USA 87: 7280-7284 (1990)), can inhibit the transforming activity of v-erb and v-fms (Lammers et al., J. Biol. Chem. 268: 22456-22462 (1993), Zander et al., Oncogene 8: 1175-1182 (1993)). Moreover, it was found that transformation by the oncogenic form of the HER2/neu gene was suppressed in NIH 3T3 fribroblasts overexpressing PTP1B (Brown-Shimer et al., Cancer Res. 52: 478-482 (1992)).
The expression level of PTP1B was found to be increased in a mammary cell line transformed with neu (Zhay et al., Cancer Res. 53: 2272-2278 (1993)). The intimate relationship between tyrosine kinases and PTPases in the development of cancer is further evidenced by the finding that PTPxcex5 is highly expressed in murine mammary tumors in transgenic mice over-expressing c-neu and v-Ha-ras, but not c-myc or int-2 (Elson and Leder, J. Biol. Chem. 270: 26116-26122 (1995)). Further, the human gene encoding PTPxcex3 was mapped to 3p21, a chromosomal region, which is frequently deleted in renal and lung carcinomas (LaForgia et al., Proc. Natl. Acad. Sci. USA 88: 5036-5040 (1991)).
In this context, it seems significant that PTPases appear to be involved in controlling the growth of fibroblasts. Thus, it was found that Swiss 3T3 cells harvested at high density contain a membrane-associated PTPase whose activity on an average is 8-fold higher than that of cells harvested at low or medium density (Pallen and Tong, Proc. Natl. Acad. Sci. USA 88: 6996-7000 (1991)). It was hypothesized by the authors that density-dependent inhibition of cell growth involves the regulated elevation of the activity of the PTPase(s) in question. In accordance with this view, a membrane-bound, receptor-type PTPase, DEP-1, showed enhanced ( greater than =10-fold) expression levels with increasing cell density of WI-38 human embryonic lung fibroblasts and in the AG1518 fibroblast cell line (xc3x96stman et al., Proc. Natl. Acad. Sci. USA 91: 9680-9684 (1994)).
Two closely related receptor-type PTPases, PTPxcexa and PTPxcexc, can mediate homophilic cell-cell interaction when expressed in non-adherent insect cells, suggesting that these PTPases might have a normal physiological function in cell-to-cell signalling (Gebbink et al., J. Biol. Chem. 268: 16101-16104 (1993), Brady-Kalnay et al., J. Cell Biol. 122: 961-972 (1993); Sap et al., Mol. Cell. Biol. 14: 1-9 (1994)). Interestingly, PTPxcexa and PTPxcexc do not interact with each other, despite their structural similarity (Zondag et al., J. Biol. Chem. 270: 14247-14250 (1995)). From the studies described above it is apparent that PTPases may play an important role in regulating normal cell growth. However, as pointed out above, other studies indicate that PTPases may also function as positive mediators of intracellular signaling and thereby induce or enhance mitogenic responses. Increased activity of certain PTPases might therefore result in cellular transformation and tumor formation. Indeed, in one study over-expression of PTPxcex1 was found to lead to transformation of rat embryo fibroblasts (Zheng, supra). In addition, SAP-1 was found to be highly expressed in pancreatic and colorectal cancer cells. SAP-1 is mapped to chromosome 19 region q13.4 and might be related to carcinoembryonic antigen mapped to 19q13.2 (Uchida et al., J. Biol. Chem. 269: 12220-12228 (1994)). Further, the dsPTPase, cdc25, dephosphorylates cdc2 at Thr14/Tyr-15 and thereby functions as positive regulator of mitosis (reviewed by Hunter, Cell 80: 225-236 (1995)). Inhibitors of specific PTPases are therefore likely to be of significant therapeutic value in the treatment of certain forms of cancer.
PTPases: Platelet Aggregation
PTPases seem to be centrally involved in platelet aggregation. Thus, agonist-induced platelet activation results in calpain-catalyzed cleavage of PTP1B with a concomitant 2-fold stimulation of PTPase activity (Frangioni et al., EMBO J. 12: 4843-4856 (1993)). The cleavage of PTP1B leads to subcellular relocation of the enzyme and correlates with the transition from reversible to irreversible platelet aggregation in platelet-rich plasma. In addition, the SH2 domain containing PTPase, SHP-1, was found to translocate to the cytoskeleton in platelets after thrombin stimulation in an aggregation-dependent manner (Li et al., FEBS Lett. 343: 89-93 (1994)).
Although some details in the above two studies have been questioned, there is over-all agreement that PTP1B and SHP-1 play significant functional roles in platelet aggregation (Ezumi et al., J. Biol. Chem. 270: 11927-11934 (1995)). In accordance with these observations, treatment of platelets with the PTPase inhibitor pervanadate leads to significant increase in tyrosine phosphorylation, secretion and aggregation (Pumiglia et al., Biochem. J. 286: 441-449 (1992)).
PTPases: Osteoporosis
The rate of bone formation is determined by the number and the activity of osteoblasts, which in term are determined by the rate of proliferation and differentiation of osteoblast progenitor cells, respectively. Histomorphometric studies indicate that the osteoblast number is the primary determinant of the rate of bone formation in humans (Gruber et al., Mineral Electrolyte Metab. 12: 246-254 (1987), reviewed in Lau et al., Biochem. J. 257: 23-36 (1989)). Acid phosphatases/PTPases may be involved in negative regulation of osteoblast proliferation. Thus, fluoride, which has phosphatase inhibitory activity, has been found to increase spinal bone density in osteoporotics by increasing osteoblast proliferation (Lau et al., supra). Consistent with this observation, an osteoblastic acid phosphatase with PTPase activity was found to be highly sensitive to mitogenic concentrations of fluoride (Lau et al., J. Biol. Chem. 260: 4653-4660 (1985), Lau et al., J. Biol. Chem. 262: 1389-1397 (1987), Lau et al., Adv. Protein Phosphatases 4: 165-198 (1987)). Interestingly, the level of membrane-bound PTPase activity was increased dramatically when the osteoblast-like cell line UMR 106.06 was grown on collagen type-I matrix compared to uncoated tissue culture plates. Since a significant increase in PTPase activity was observed in density-dependent growth arrested fibroblasts (Pallen and Tong, Proc. Natl. Acad. Sci. 88: 6996-7000 (1991)), it might be speculated that the increased PTPase activity directly inhibits cell growth. The mitogenic action of fluoride and other phosphatase inhibitors (molybdate and vanadate) may thus be explained by their inhibition of acid phosphatases/PTPases that negatively regulate the cell proliferation of osteoblasts. The complex nature of the involvement of PTPases in bone formation is further suggested by the identification of a novel parathyroid regulated, receptor-like PTPase, OST-PTP, expressed in bone and testis (Mauro et al., J. Biol. Chem. 269: 30659-30667 (1994)). OST-PTP is up-regulated following differentiation and matrix formation of primary osteoblasts and subsequently down-regulated in the osteoblasts which are actively mineralizing bone in culture. It may be hypothesized that PTPase inhibitors may prevent differentiation via inhibition of OST-PTP or other PTPases thereby leading to continued proliferation. This would be in agreement with the above-mentioned effects of fluoride and the observation that the tyrosine phosphatase inhibitor orthovanadate appears to enhance osteoblast proliferation and matrix formation (Lau et al., Endocrinology 116: 2463-2468 (1988)). In addition, it was observed that vanadate, vanadyl and pervanadate all increased the growth of the osteoblast-like cell line UMR106. Vanadyl and pervanadate were stronger stimulators of cell growth than vanadate. Only vanadate was able to regulate the cell differentiation as measured by cell alkaline phosphatase activity (Cortizo et al., Mol. Cell. Biochem. 145: 97-102 (1995)). It is of particular interest to the current invention that several studies have shown that bisphosphonates, such as alendronate and tiludronate, inhibit the PTPase activity in osteoclasts, and that the inhibition of PTPase activity correlated with the inhibition of in vitro osteoclast formation and boneresorption (Schmidt et al., Proc. Natl. Acad. Sci. U.S.A. 93: 3068-3073 (1996); Murakami et al., Bone 20: 399-404 (1997); Opas et al., Biochem. Pharmacol. 54: 721-727 (1997); Skorey et al., J. Biol. Chem. 272: 22472-22480 (1997)). Thus, PTPase inhibitorsxe2x80x94other than bisphophonatesxe2x80x94can potentially be effective for prevention and/or treatment of osteoporosis.
PTPases: Microorganisms
Dixon and coworkers have called attention to the fact that PTPases may be a key element in the pathogenic properties of Yersinia (reviewed in Clemens et al. Molecular Microbiology 5: 2617-2620 (1991)). This finding was rather surprising since tyrosine phosphate is thought to be absent in bacteria. The genus Yersinia comprises 3 species: Y. pestis (responsible for the bubonic plague), Y. pseudoturberculosis and Y. enterocolitica (causing enteritis and mesenteric lymphadenitis). Interestingly, a dual-specificity phosphatase, VH1, has been identified in Vaccinia virus (Guan et al., Nature 350: 359-263 (1991)). These observations indicate that PTPases may play critical roles in microbial and parasitic infections, and they further point to PTPase inhibitors as a novel, putative treatment principle of infectious diseases.
WO 99/46267 discloses compounds which are pharmacologically useful inhibitors of PTPases. However, the present invention which represents a novel selection under WO 99/46267, discloses a class of compounds which surprisingly are more potent against protein tyrosine phosphatases (e.g. PTP1B) than those disclosed in WO 99/46267.
The present invention relates to Compounds of the Formula 1 wherein n, m, X, R1, R2, R3, R4, R5 and R6 are defined below; 
In the above Formula 1
n is 0, 1 or 2;
m is 1 or 2;
X is S or O;
R1 is hydrogen, COOR3, or R1 is selected from the group consisting of the following 5-membered heterocycles: 
R2 is hydrogen, C1-C6alkyl, hydroxy or NR8R9;
R3 is hydrogen, C1-C6alkyl, arylC1-C6alkyl, C1-C6alkylcarbonyloxyC1-C6alkyl or C1-C6alkylcarbonyloxyarylC1-C6alkyl;
R4, R5 and R6 are independently hydrogen, trihalomethyl, C1-C6alkyl, aryl, arylC1-C6alkyl, hydroxy, oxo, carboxy, carboxyC1-C6alkyl, C1-C6alkyloxy-carbonyl, aryloxycarbonyl, arylC1-C6alkyloxycarbonyl, C1-C6alkyloxy, C1-C6alkyloxyC1-C6alkyl, aryloxy, arylC1-C6alkyloxy, arylC1-C6alkyloxyC1-C6alkyl, thio, C1-C6alkylthio, C1-C6alkylthioC1-C6alkyl, arylthio, arylC1-C6alkylthio, arylC1-C6alkylthioC1-C6alkyl, NR8R9, R8R9NC1-C6alkyl, C1-C6alkylaminoC1-C6alkyl, arylaminoC1-C6alkyl, arylC1-C6alkylaminoC1-C6alkyl, di(arylC1-C6aminoC1-C6alkyl, C1-C6alkylcarbonyl, C1-C6alkyl-carbonylC1-C6alkyl, arylC1-C6alkylcarbonyl, arylC1-C6alkylcarbonylC1-C6alkyl, C1-C6alkylcarboxy, C1-C6alkylcarboxyC1-C6-alkyl, arylcarboxy, arylcarboxyC1-C6alkyl, arylC1-C6alkylcarboxy, arylC1-C6alkylcarboxyC1-C6alkyl, C1-C6alkylcarbonylamino, C1-C6alkylcarbonylaminoC1-C6alkyl, arylcarbonylaminoC1-C6alkyl, -carbonyINR8C1-C6alkylCOR12, arylC1-C6alkylcarbonylamino, arylC1-C6alkylcarbonylaminoC1-C6alkyl, CONR8R9, or C1-C6alkylCONR8R9 wherein the alkyl and aryl groups are optionally substituted and R12 is NR8R9, or C1-C6alkylNR8R9;
R7 is hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, C1-C6alkoxycarbonyl, arylcarbonyl, aryloxocarbonyl, arylC1-C6alkylcarbonyl, arylC1-C6alkoxycarbonyl, C1-C6alkylcarboxy, arylC1-C6alkylcarboxy, R10R11NcarbonylC1-C6alkyl wherein R10 and R11 are independently selected from hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, arylcarbonyl, arylC1-C6alkylcarbonyl, C1-C6alkylcarboxy or arylC1-C6alkyl-carboxy; wherein the alkyl and aryl groups are optionally substituted;
R8 and R9 are independently selected from hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, C1-C6alkoxycarbonyl, arylcarbonyl, aryloxocarbonyl, arylC1-C6alkylcarbonyl, arylC1-C6alkoxycarbonyl, C1-C6alkylcarboxy, arylC1-C6alkylcarboxy, R10R11NcarbonylC1-C6alkyl wherein the alkyl and aryl groups are optionally substituted; or R8 and R9 are together with the nitrogen to which they are attached forming a saturated, partially saturated or aromatic cyclic, bicyclic or tricyclic ring system containing from 3 to 14 carbon atoms and from 0 to 3 additional heteroatoms selected from nitrogen, oxygen or sulphur, the ring system can optionally be substituted with at least one C1-C6alkyl, aryl, arylC1-C6alkyl, hydroxy, oxo, C1-C6alkyloxy, arylC1-C6alkyloxy, C1-C6alkyloxyC1-C6alkyl, NR10R11 or C1-C6alkylaminoC1-C6alkyl, wherein R10 and R11 are independently selected from hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, arylcarbonyl, arylC1-C6alkylcarbonyl, C1-C6alkylcarboxy or arylC1-C6alkylcarboxy; wherein the alkyl and aryl groups are optionally substituted; or
R8 and R9 are independently a saturated or partial saturated cyclic 5, 6 or 7 membered amine, imide or lactam;
or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric form, or prodrug thereof.
The compounds of the invention can be further modified to act as prodrugs.
It is a well known problem in drug discovery that compounds, such as enzyme inhibitors, may be very potent and selective in biochemical assays, yet be inactive in vivo. This lack of so-called bioavailability may be ascribed to a number of different factors such as lack of or poor absorption in the gut, first pass metabolism in the liver, poor uptake in cells. Although the factors determining bioavailability are not completely understood, there are many examples in the scientific literaturexe2x80x94well known to those skilled in the artxe2x80x94of how to modify compounds, which are potent and selective in biochemical assays but show low or no activity in vivo, into drugs that are biologically active. It is within the scope of the invention to modify the compounds of the invention, termed the xe2x80x98original compoundxe2x80x99, by attaching chemical groups that will improve the bioavailability of said compounds in such a way that the uptake in cells or mammals is facilitated. Examples of said modifications, which are not intended in any way to limit the scope of the invention, include changing of one or more carboxy groups to esters (for instance methyl esters, ethyl esters, acetoxymethyl esters or other acyloxymethyl esters). Compounds of the invention, original compounds, such modified by attaching chemical groups are termed xe2x80x98modified compoundsxe2x80x99. Said chemical groups may or may not be apparent in the claims of this invention. Other examples of modified compounds, which are not intended in any way to limit the scope of the invention, are compounds that have been cyclized at specific positionsxe2x80x94socalled xe2x80x98cyclic compoundsxe2x80x99xe2x80x94which upon uptake in cells or mammals become hydrolyzed at the same specific position(s) in the molecule to yield the compounds of the invention, the original compounds, which are then said to be xe2x80x98non-cyclicxe2x80x99. For the avoidance of doubt, it is understood that the latter original compounds in most cases will contain other cyclic or heterocyclic structures that will not be hydrolyzed after uptake in cells or mammals. Generally, said modified compounds will not show a behaviour in bio-chemical assays similar to that of the original compound, i.e. the corresponding compounds of the invention without the attached chemical groups or said modifications. Said modified compounds may even be inactive in biochemical assays. However, after uptake in cells or mammals these attached chemical groups of the modified compounds may in turn be removed spontaneously or by endogenous enzymes or enzyme systems to yield compounds of the invention, original compounds. xe2x80x98Uptakexe2x80x99 is defined as any process that will lead to a substantial concentration of the compound inside cells or in mammals. After uptake in cells or mammals and after removal of said attached chemical group or hydrolysis of said cyclic compound, the compounds may have the same structure as the original compounds and thereby regain their activity and hence become active in cells and/or in vivo after uptake. A number of procedures, well known to those skilled in the art, may be used to verify that the attached chemical groups have been removed or that the cyclic compound has been hydrolyzed after uptake in cells or mammals. An example, which is not intended in any way to limit the scope of the invention, is given in the following. A mammalian cell line, which can be obtained from the American Tissue Type Collection or other similar governmental or commercial sources, is incubated with said modified compound. After incubation at conditions well known to those skilled in the art, the cells are washed appropriately, lysed and the lysate is isolated. Appropriate controls, well known to those skilled in the art, must be included. A number of different procedures, well known to those skilled in the art, may in turn be used to extract and purify said compound from said lysate. Said compound may or may not retain the attached chemical group or said cyclic compound may or may not have been hydrolyzed. Similarly, a number of different proceduresxe2x80x94well known to those skilled in the artxe2x80x94may be used to structurally and chemically characterize said purified compound. Since said purified compound has been isolated from said cell lysate and hence has been taken up by said cell line, a comparison of said structurally and chemically characterized compound with that of the original unmodified compound (i.e. without said attached chemical group or said non-cyclic compound) will immediately provide those skilled in the art information on whether the attached chemical group as been removed in the cell or if the cyclic compound has been hydrolyzed. As a further analysis, said purified compound may be subjected to enzyme kinetic analysis as described in detail in the present invention. If the kinetic profile is similar to that of the original compound without said attached chemical group, but different from said modified compound, this confirms that said chemical group has been removed or said cyclic compounds has been hydrolyzed. Similar techniques may be used to analyze compounds of the invention in whole animals and mammals.
A preferred prodrug is acetoxymethyl esters of the compounds of the present invention which may be prepared by the following general procedure (C. Schultz et al, The Journal of Biological Chemistry, 1993, 268, 6316-6322.):
A carboxylic acid (1 equivalent) is suspended in dry acetonitrile (2 ml per 0.1 mmol). Diisopropyl amine (3.0 equivalents) is added followed by bromomethyl acetate (1.5 equivalents). The mixture is stirred under nitrogen overnight at room temperature. Acetonitrile is removed under reduced pressure to yield an oil which is diluted in ethylacetate and washed with water (3xc3x97). The organic layer is dried over anhydrous magnesium sulfate. Filtration followed by solvent removal under reduced pressure afford a crude oil. The product is purified by column chromatography on silica gel, using an appropriate solvent system.
As used herein, the term xe2x80x9cattachedxe2x80x9d or xe2x80x9cxe2x80x94xe2x80x9d (e.g. xe2x80x94COR11 which indicates the carbonyl attachment point to the scaffold) signifies a stable covalent bond, certain preferred points of attachment points being apparent to those skilled in the art.
The terms xe2x80x9chalogenxe2x80x9d or xe2x80x9chaloxe2x80x9d include fluorine, chlorine, bromine, and iodine.
The term xe2x80x9calkylxe2x80x9d includes C1-C6 straight chain saturated, methylene and C2-C6 unsaturated aliphatic hydrocarbon groups, C1-C6 branched saturated and C2-C6 unsaturated aliphatic hydrocarbon groups, C3-C6 cyclic saturated and C5-C6 unsaturated aliphatic hydrocarbon groups, and C1-C6 straight chain or branched saturated and C2-C6 straight chain or branched unsaturated aliphatic hydrocarbon groups substituted with C3-C6 cyclic saturated and unsaturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, this definition shall include but is not limited to methyl (Me), ethyl (Et), propyl (Pr), butyl (Bu), pentyl, hexyl, heptyl, ethenyl, propenyl, butenyl, penentyl, hexenyl, isopropyl (i-Pr), isobutyl (i-Bu), tert-butyl (t-Bu), sec-butyl (s-Bu), isopentyl, neopentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl, cyclohexenyl, methylcyclopropyl, ethylcyclohexenyl, butenylcyclopentyl, and the like.
The term xe2x80x9csubstituted alkylxe2x80x9d or xe2x80x9coptionally substituted alkylxe2x80x9d represents an alkyl group as defined above wherein the substitutents are independently selected from halo, cyano, nitro, trihalomethyl, carbamoyl, hydroxy, oxo, COOR3, CONR8R9, C1-C6alkyl, C1-C6alkyloxy, aryloxy, arylC1-C6alkyloxy, thio, C1-C6alkylthio, arylthio, arylC1-C6alkylthio, NR8R9, C1-C6alkylamino, arylamino, arylC1-C6alkylamino, di(arylC1-C6alkyl)amino, C1-C6alkylcarbonyl, arylC1-C6-alkylcarbonyl, C1-C6alkylcarboxy, arylcarboxy, arylC1-C6alkylcarboxy, C1-C6alkylcarbonylamino, -C1-C6alkylaminoCOR12, arylC1-C6alkylcarbonylamino, tetrahydrofuranyl, morpholinyl, piperazinyl, xe2x80x94CONR8R9, xe2x80x94C1-C6alkylCONR8R9, or a saturated or partial saturated cyclic 5, 6 or 7 membered amine, imide or lactam; wherein R11 is hydroxy, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkyloxy, aryloxy, arylC1-C6alkyloxy and R3 is defined as above or NR8R9, wherein R8, R9 are defined as above.
The term xe2x80x9csaturated, partially saturated or aromatic cyclic, bicyclic or tricyclic ring systemxe2x80x9d represents but are not limit to aziridinyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, imidazolyl, 2-imidazolinyl, imidazolidinyl, pyrazolyl, 2-pyrazolinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, morpholinyl, piperidinyl, thiomorpholinyl, piperazinyl, indolyl, isoindolyl, 1,2,3,4-tetrahydro-quinolinyl, 1 ,2,3,4-tetrahydro-isoquinolinyl, 1,2,3,4-tetrahydro-quinoxalinyl, indolinyl, indazolyl, benzimidazolyl, benzotriazolyl, purinyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl, iminodibenzyl, iminostilbenyl.
The term xe2x80x9calkyloxyxe2x80x9d (e.g. methoxy, ethoxy, propyloxy, allyloxy, cyclohexyloxy) represents an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms attached through an oxygen bridge.
The term xe2x80x9calkyloxyalkylxe2x80x9d represents an xe2x80x9calkyloxyxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms.
The term xe2x80x9calkyloxyalkyloxyxe2x80x9d represents an xe2x80x9calkyloxyalkylxe2x80x9d group attached through an oxygen atom as defined above having the indicated number of carbon atoms.
The term xe2x80x9caryloxyxe2x80x9d (e.g. phenoxy, naphthyloxy and the like) represents an aryl group as defined below attached through an oxygen bridge.
The term xe2x80x9carylalkyloxyxe2x80x9d (e.g. phenethyloxy, naphthylmethyloxy and the like) represents an xe2x80x9carylalkylxe2x80x9d group as defined below attached through an oxygen bridge.
The term xe2x80x9carylalkyloxyalkylxe2x80x9d represents an xe2x80x9carylalkyloxyxe2x80x9d group as defined above attached through an xe2x80x9calkylxe2x80x9d group defined above having the indicated number of carbon atoms.
The term xe2x80x9carylthioxe2x80x9d (e.g. phenylthio, naphthylthio and the like) represents an xe2x80x9carylxe2x80x9d group as defined below attached through an sulfur bridge.
The term xe2x80x9calkyloxycarbonylxe2x80x9d (e.g. methylformiat, ethylformiat and the like) represents an xe2x80x9calkyloxyxe2x80x9d group as defined above attached through a carbonyl group.
The term xe2x80x9caryloxycarbonylxe2x80x9d (e.g. phenylformiat, 2-thiazolylformiat and the like) represents an xe2x80x9caryloxyxe2x80x9d group as defined above attached through a carbonyl group.
The term xe2x80x9carylalkyloxycarbonylxe2x80x9d (e.g. benzylformiat, phenyletylformiat and the like) represents an xe2x80x9carylalkyloxyxe2x80x9d group as defined above attached through a carbonyl group.
The term xe2x80x9calkyloxycarbonylalkylxe2x80x9d represents an xe2x80x9calkyloxycarbonylxe2x80x9d group as defined above attached through an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms.
The term xe2x80x9carylalkyloxycarbonylalkylxe2x80x9d represents an xe2x80x9carylalkyloxycarbonylxe2x80x9d group as defined above attached through an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms.
The term xe2x80x9calkylthioxe2x80x9d (e.g. methylthio, ethylthio, propylthio, cyclohexenylthio and the like) represents an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms attached through a sulfur bridge.
The term xe2x80x9carylalkylthioxe2x80x9d (e.g. phenylmethylthio, phenylethylthio, and the like) represents an xe2x80x9carylalkylxe2x80x9d group as defined above having the indicated number of carbon atoms attached through a sulfur bridge.
The term xe2x80x9calkylthioalkylxe2x80x9d represents an xe2x80x9calkylthioxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms.
The term xe2x80x9carylalkylthioalkylxe2x80x9d represents an xe2x80x9carylalkylthioxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms.
The term xe2x80x9calkylaminoxe2x80x9d (e.g. methylamino, diethylamino, butylamino, N-propyl-N-hexylamino, (2-cyclopentyl)propylamino, hexenylamino, pyrrolidinyl, piperidinyl and the like) represents one or two xe2x80x9calkylxe2x80x9d groups as defined above having the indicated number of carbon atoms attached through an amine bridge. The two alkyl groups may be taken together with the nitrogen to which they are attached forming a saturated, partially saturated or aromatic cyclic, bicyclic or tricyclic ring system containing 3 to 14 carbon atoms and 0 to 3 additional heteroatoms selected from nitrogen, oxygen or sulfur, the ring system can optionally be substituted with at least one C1-C6alkyl, aryl, arylC1-C6alkyl, hydroxy, oxo, C1-6alkyloxy, C1-C6alkyloxyC1-C6alkyl, NR8R9, C1-C6alkylaminoC1-C6alkyl substituent wherein the alkyl and aryl groups are optionally substituted as defined in the definition section and R8 and R9 are defined as above.
The term xe2x80x9carylalkylaminoxe2x80x9d (e.g. benzylamino, diphenylethylamino and the like) represents one or two xe2x80x9carylalkylxe2x80x9d groups as defined above having the indicated number of carbon atoms attached through an amine bridge. The two xe2x80x9carylalkylxe2x80x9d groups may be taken together with the nitrogen to which they are attached forming a saturated, partially saturated or aromatic cyclic, bicyclic or tricyclic ring system containing 3 to 14 carbon atoms and 0 to 3 additional heteroatoms selected from nitrogen, oxygen or sulfur, the ring system can optionally be substituted with at least one C1-C6alkyl, aryl, arylC1-C6alkyl, hydroxy, oxo, C1-C6alkyloxy, C1-C6alkyloxyC1-C6alkyl, NR8R9, C1-C6alkylaminoC1-C6alkyl substituent wherein the alkyl and aryl groups are optionally substituted as defined in the definition section and R8 and R9 are defined as above.
The term xe2x80x9calkylaminoalkylxe2x80x9d represents an xe2x80x9calkylaminoxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms.
The term xe2x80x9carylalkylaminoalkylxe2x80x9d represents an xe2x80x9carylalkylaminoxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms. 10 The term xe2x80x9carylaminoxe2x80x9d represents an xe2x80x9carylxe2x80x9d group as defined below attached through an amino group.
The term xe2x80x9carylaminoalkylxe2x80x9d represents an xe2x80x9carylaminoxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms.
The term xe2x80x9carylalkylxe2x80x9d (e.g. benzyl, phenylethyl) represents an xe2x80x9carylxe2x80x9d group as defined below attached through an alkyl having the indicated number of carbon atoms or substituted alkyl group as defined above.
The term xe2x80x9calkylcarbonylxe2x80x9d (e.g. cyclooctylcarbonyl, pentylcarbonyl, 3-hexenylcarbonyl) represents an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms attached through a carbonyl group.
The term xe2x80x9carylcarbonylxe2x80x9d (benzoyl) represents an xe2x80x9carylxe2x80x9d group as defined above attached through a carbonyl group.
The term xe2x80x9carylalkylcarbonylxe2x80x9d (e.g. phenylcyclopropylcarbonyl, phenylethylcarbonyl and the like) represents an xe2x80x9carylalkylxe2x80x9d group as defined above having the indicated number of carbon atoms attached through a carbonyl group.
The term xe2x80x9calkylcarbonylalkylxe2x80x9d represents an xe2x80x9calkylcarbonylxe2x80x9d group attached through an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms.
The term xe2x80x9carylalkylcarbonylalkylxe2x80x9d represents an xe2x80x9carylalkylcarbonylxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms.
The term xe2x80x9carylcarbonylaminoxe2x80x9d represents an xe2x80x9carylcarbonylxe2x80x9d group as defined above attached through an amino group.
The term xe2x80x9carylcarbonylaminoalkylxe2x80x9d represents an xe2x80x9carylcarbonylaminoxe2x80x9d group attached through an alkyl group as defined above having the indicated number of carbon atoms.
The term xe2x80x9calkylcarboxyxe2x80x9d (e.g. heptylcarboxy, cyclopropylcarboxy, 3-pentenylcarboxy) represents an xe2x80x9calkylcarbonylxe2x80x9d group as defined above wherein the carbonyl is in turn attached through an oxygen bridge.
The term xe2x80x9carylcarboxyalkylxe2x80x9d (e.g. phenylcarboxymethyl) represents an xe2x80x9carylcarbonylxe2x80x9d group defined above wherein the carbonyl is in turn attached through an oxygen bridge to an alkyl chain having the indicated number of carbon atoms.
The term xe2x80x9carylalkylcarboxyxe2x80x9d (e.g. benzylcarboxy, phenylcyclopropylcarboxy and the like) represents an xe2x80x9carylalkylcarbonylxe2x80x9d group as defined above wherein the carbonyl is in turn attached through an oxygen bridge.
The term xe2x80x9calkylcarboxyalkylxe2x80x9d represents an xe2x80x9calkylcarboxyxe2x80x9d group attached through an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms.
The term xe2x80x9carylalkylcarboxyalkylxe2x80x9d represents an xe2x80x9carylalkylcarboxyxe2x80x9d group attached through an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms.
The term xe2x80x9calkylcarbonylaminoxe2x80x9d (e.g. hexylcarbonylamino, cyclopentylcarbonyl- aminomethyl, methylcarbonylaminophenyl) represents an xe2x80x9calkylcarbonylxe2x80x9d group as defined above wherein the carbonyl is in turn attached through the nitrogen atom of an amino group. The nitrogen atom may itself be substituted with an alkyl or aryl group.
The term xe2x80x9carylalkylcarbonylaminoxe2x80x9d (e.g. benzylcarbonylamino and the like) represents an xe2x80x9carylalkylcarbonylxe2x80x9d group as defined above wherein the carbonyl is in turn attached through the nitrogen atom of an amino group. The nitrogen atom may itself be substituted with an alkyl or aryl group.
The term xe2x80x9calkylcarbonylaminoalkylxe2x80x9d represents an xe2x80x9calkylcarbonylaminoxe2x80x9d group attached through an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms. The nitrogen atom may itself be substituted with an alkyl or aryl group.
The term xe2x80x9carylalkylcarbonylaminoalkylxe2x80x9d represents an xe2x80x9carylalkylcarbonylaminoxe2x80x9d group attached through an xe2x80x9calkylxe2x80x9d group as defined above having the indicated number of carbon atoms. The nitrogen atom may itself be substituted with an alkyl or aryl group.
The term xe2x80x9calkylcarbonylaminoalkylcarbonylxe2x80x9d represents an alkylcarbonylaminoalkyl group attached through a carbonyl group. The nitrogen atom may be further substituted with an xe2x80x9calkylxe2x80x9d or xe2x80x9carylxe2x80x9d group.
The term xe2x80x9carylxe2x80x9d represents an unsubstituted, monocyclic, polycyclic, biaryl and heterocyclic aromatic groups covalently attached at any ring position capable of forming a stable covalent bond, certain preferred points of attachment being apparent to those skilled in the art (e.g., 3-indolyl, 4(5)-imidazolyl).
The definition of aryl includes but is not limited to phenyl, biphenyl, indenyl, fluorenyl, naphthyl (1-naphthyl, 2-naphthyl), pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1,2,3-triazol-1-yl, 1,2,3-triazol-2-yl 1,2,3-triazol4-yl, 1,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), isoxazolyl (3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl), thiazolyl (2-thiazolyl, 4-thiazolyl, 5-thiazolyl), thiophenyl (2-thiophenyl, 3-thiophenyl, 4-thiophenyl, 5-thiophenyl), furanyl (2-furanyl, 3-furanyl, 4-furanyl, 5-furanyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl), 5-tetrazolyl, pyrimidiny (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl), quinolyl (2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl (1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5-isoquinolyl, 6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b]furanyl, 5-benzo[b]furanyl, 6-benzo[b]furanyl, 7-benzo[b]furanyl), 2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro-benzo[b]furanyl), 3-(2,3-dihydro-benzo[b]furanyl), 4-(2,3-dihydro-benzo[b]furanyl), 5-(2,3-dihydro-benzo-[b]furanyl), 6-(2,3-dihydro-benzo-[b]furanyl) 7-(2,3-dihydro-benzo[b]furanyl)), benzo[b]thiophenyl (2-benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl, 5-benzo[b]thiophenyl, 6-benzo[b]thiophenyl, 7-benzo[b]thiophenyl), 2,3-dihydro-benzo[b]-thiophenyl (2-(2,3-dihydro-benzo[b]thiophenyl), 3-(2,3-dihydro-benzo[b]-thiophenyl), 4-(2,3-dihydro-benzo[b]thiophenyl), 5-(2,3-dihydro-benzo[b ]-thiophenyl), 6-(2,3-dihydro-benzo[b]thiophenyl), 7-(2, 3-dihydro-benzo[b]-thiophenyl)), 4,5,6,7-tetrahydro-benzo[b]thiophenyl (2-(4,-5,6,7=tetrahydro-benzo-[b]thiophenyl), 3-(4,5,6,7-tetrahydro-benzo-[b]thiophenyl), 4-(4,5,6,7-tetrahydro-benzo[b]thiophenyl), 5-(4,5,6,7-tetrahydro-benzo-[b]thiophenyl), 6-(4, 5,6,7tetrahydro-benzo-[b]thiophenyl), 7-(4,5,6,7-tetrahydro-benzo[b]thiophenyl)), 4-,5,6,7-tetrahydro-thieno[2,3-c]pyridyl (4-(4,5,6,7-tetrahydro-thieno[2,3-c]pyridyl), 5-4,5,6,7-tetrahydro-thieno[2,3-c]pyridyl) 6-(4,5,6,7-tetrahydro-thieno[2,3-c]pyridyl), 7-(4,5,6,7-tetrahydro-thieno[2,3-c]pyridyl)), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), isoindolyl (1-isoindolyl, 2-isoindolyl, 3-isoindolyl, 4-isoindolyl, 5-isoindolyl, 6-isoindolyl, 7-isoindolyl), 1,3-dihydro -isoindolyl (1-(1,3-dihydro-isoindolyl), 2-(1,3-dihydro-isoindolyl), 3-(1,3-dihydro-isoindolyl), 4-(1,3-dihydro-isoindolyl), 5-(1,3-dihydro-isoindolyl), 6-(1,3-dihydro-isoindolyl), 7-(1,3-dihydro -isoindolyl)), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzo-oxazolyl, 2-benzoxazolyl), benzothiazolyl (1-benzothiazolyl, 2-benzo-thiazolyl, 4-benzothiazolyl, 5-benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl), 5H-dibenz[b,f]azepine (5H-dibenz[b,f]azepin-1-yl, 5H-dibenz-[b,f]azepine-2-yl, 5H-dibenz[b,f]azepine-3-yl, 5H-dibenz-[b,f]azepine4-yl, 5H-dibenz[b,f]-azepine-5-yl), 10,11-dihydro-5H-dibenz[b,f]azepine (10,11 -dihydro-5H-dibenz[b,f]azepine-1-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-2-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-3-yl, 10,11-dihydro-5H-dibenz-[b,f]azepine-4-yl, 10,11-dihydro-5H-dibenz [b,f]azepine-5-yl), piperidinyl (2-piperidinyl, 3-piperidinyl, 4-piperidinyl), pyrrolidinyl (1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl), phenylpyridyl (2-phenyl-pyridyl, 3-phenyl-pyridyl, 4-phenylpridyl), phenylpyrimidinyl (2-phenylpyrimidinyl, 4-phenyl-pyrimidinyl, 5-phenylpyrimidinyl, 6-phenylpyrimidinyl), phenylpyrazinyl, phenylpyridazinyl (3-phenylpyridazinyl, 4-phenylpyridazinyl, 5-phenyl-pyridazinyl).
The term xe2x80x9coptionally substituted arylxe2x80x9d represents an mono-, di- or trisubstituted aryl as defined above wherein the substituents are independently selected from the group consisting of halo, nitro, cyano, trihalomethyl, C1-C6alkyl, aryl, arylC1-C6alkyl, hydroxy, COOR3, CONR8R9, C1-C 6alkyloxy, C1-C6alkyloxyC1-C6alkyl, aryloxy, arylC1-C6alkyloxy, arylC1-C6alkyloxyC1-C6alkyl, thio, C1-C6alkylthio, C1-C6alkylthioC1-C6alkyl, arylthio, arylC1-C6alkylthio, arylC1-C6alkylthioC1-C6alkyl, NR8R9, C1-C6-alkylamino, C1-C6(alkyl-aminoC1-C6alkyl, arylamino, arylC1-C6alkylamino, arylC1-C6alkyl-aminoC1-C6alkyl, di(arylC1-C6alkyl)aminoC1-C6alkyl, C1-C6alkylcarbonyl, C1-C6alkyl-carbonylC1-C6alkyl, arylC1-C6alkylcarbonyl, arylC1-C6alkylcarbonylC1-C6alkyl, C1-C6alkyl-carboxy, C1-C6alkylcarboxyC1-C6alkyl, arylC1-C6alkylcarboxy, arylC1-C6alkyl-carboxyC1-C6alkyl, carboxyC1-C6-alkyloxy, C1-C6alkylcarbonylamino, C1-C6alkyl-carbonylaminoC1-C6alkyl, -carbonyINR7C1-C6alkylCOR11, arylC1-C6alkylcarbonyl-amino, arylC1-C6-alkylcarbonylaminoC1-C6alkyl, xe2x80x94CONR8R9, or xe2x80x94C1-C6alkylCONR8R9; wherein R3, R8, R9, and R11, are defined as above and the alkyl and aryl groups are optionally substituted as defined in the definition section;
The term xe2x80x9carylcarbonylxe2x80x9d (e.g. 2-thiophenylcarbonyl, 3-methoxy-anthrylcarbonyl, oxazolylcarbonyl) represents an xe2x80x9carylxe2x80x9d group as defined above attached through a carbonyl group.
The term xe2x80x9carylalkylcarbonylxe2x80x9d (e.g. (2,3-dimethoxyphenyl)propylcarbonyl, (2-chloronaphthyl)pentenylcarbonyl, imidazolylcyclopentylcarbonyl) represents an xe2x80x9carylalkylxe2x80x9d group as defined above wherein the xe2x80x9calkylxe2x80x9d group is in turn attached through a carbonyl.
The compounds of the present invention have asymmetric centers and may occur as racemates, racemic mixtures, and as individual enantiomers or diastereoisomers, with all isomeric forms being included in the present invention as well as mixtures thereof.
Pharmaceutically acceptable salts of the Compounds of Formula 1, where a basic or acidic group is present in the structure, are also included within the scope of this invention. When an acidic substituent is present, such as xe2x80x94COOH, 5-tetrazolyl or xe2x80x94P(O)(OH)2, there can be formed the ammonium, morpholinium, sodium, potassium, barium, calcium salt, and the like, for use as the dosage form. When a basic group is present, such as amino or a basic heteroaryl radical, such as pyridyl, an acidic salt, such as hydrochloride, hydrobromide, phosphate, sulfate, trifluoroacetate, trichloroacetate, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartarate, fumarate, mandelate, benzoate, cinnamate, methanesulfonate, ethane sulfonate, picrate and the like, and include acids related to the pharmaceutically acceptable salts listed in Journal of Pharmaceutical Science, 66, 2 (1977) and incorporated herein by reference, can be used as the dosage form.
Also, in the case of the xe2x80x94COOH or xe2x80x94P(O)(OH)2 being present, pharmaceutically acceptable esters can be employed, e.g., methyl, tert-butyl, pivaloyloxymethyl, and the like, and those esters known in the art for modifying solubility or hydrolysis characteristics for use as sustained release or prodrug formulations.
In addition, some of the compounds of the present invention may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention.
The term xe2x80x9ctherapeutically effective amountxe2x80x9d shall mean that amount of drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor or other. In a preferred embodiment, the present invention is concerned with compounds of Formula I 
wherein
n is 0, 1 or 2;
m is 1 or 2;
X is S or O;
R1 is hydrogen or COOR3, or R1 is selected from the group consisting of the following 5-membered heterocycles: 
R2 is hydrogen, C1-C6alkyl, hydroxy or NR8R9;
R3 is hydrogen, C1-C6alkyl, arylC1-C6alkyl, C1-C6alkylcarbonyloxyC1-C6alkyl or C1-C6alkylcarbonyloxyarylC1-C6alkyl;
R4, R5 and R6 are independently hydrogen, trihalomethyl, C1-C6alkyl, aryl, arylC1-C6alkyl, hydroxy, oxo, carboxy, carboxyC1-C6alkyl, C1-C6alkyloxy-carbonyl, aryloxycarbonyl, arylC1-C6alkyloxycarbonyl, C1-C6alkyloxy, C1-C6alkyloxyC1-C6alkyl, aryloxy, arylC1-C6alkyloxy, arylC1-C6alkyloxyC1-C6alkyl, thio, C1-C6alkyl-thio, C1-C6alkylthioC1-C6alkyl, arylthio, arylC1-C6alkylthio, arylC1-C6alkylthioC1-C6alkyl, NR8R9, C1-C6alkylaminoC1-C6alkyl, arylC1-C6alkylaminoC1-C6alkyl, di(arylC1-C6alkyl)aminoC1-C6alkyl, C1-C6alkylcarbonyl, C1-C6alkylcarbonylC1-C6alkyl, arylC1-C6alkylcarbonyl, arylC1-C6alkylcarbonylC1-C6alkyl, C1-C6alkyl-carboxy, C1-C6alkylcarboxyC1-C6-alkyl, arylcarboxy, arylcarboxyC1-C6alkyl, C1-C6alkylcarboxy, arylC1-C6alkylcarboxyC1-C6alkyl, C1-C6alkylcarbonylamino, C1-C6alkylcarbonylaminoC1-C6alkyl, -carbonylNR8C1-C6alkylCOR12, arylC1-C6alkylcarbonylamino, arylC1-C6alkylcarbonylaminoC1-C6alkyl, CONR8R9, or C1-C6alkylCONR8R9 wherein the alkyl and aryl groups are optionally substituted and R12 is NR8R9, or C1-C6alkynyl8R9;
R7 is hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, C1-C6alkoxycarbonyl, arylcarbonyl, aryloxocarbonyl, arylC1-C6alkylcarbonyl, arylC1-C6alkoxycarbonyl, C1-C6alkylcarboxy, arylC1-C6alkylcarboxy, R10R11NcarbonylC1-C6alkyl wherein R10 and R11 are independently selected from hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, arylcarbonyl, arylC1-C6alkylcarbonyl, C1-C6alkylcarboxy or arylC1-C6alkylcarboxy; wherein the alkyl and aryl groups are optionally substituted; R8 and R9 are independently selected from hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, C1-C6alkoxycarbonyl, arylcarbonyl, aryloxocarbonyl, arylC1-C6alkylcarbonyl, arylC1-C6alkoxycarbonyl, C1-C6alkylcarboxy, arylC1-C6alkylcarboxy, R10R11NcarbonylCl-C6alkyl wherein the alkyl and aryl groups are optionally substituted; or
R8 and R9 are together with the nitrogen to which they are attached forming a saturated, partially saturated or aromatic cyclic, bicyclic or tricyclic ring system containing from 3 to 14 carbon atoms and from 0 to 3 additional heteroatoms selected from nitrogen, oxygen or sulphur, the ring system can optionally be substituted with at least one C1-C6alkyl, aryl, arylC1-C6alkyl, hydroxy, oxo, C1-6alkyloxy, arylC1-C6alkyloxy, C1-C6alkyloxyC1-C6alkyl, NR10R11 or C1-C6alkylaminoC1-C6alkyl, wherein R10 and R11 are independently selected from hydrogen, C1-C6alkyl, aryl, arylC1-C6alkyl, C1-C6alkylcarbonyl, arylcarbonyl, arylC1-C6alkylcarbonyl, C1-C6alkylcarboxy or arylC1-C6alkylcarboxy; wherein the alkyl and aryl groups are optionally substituted; or
R8 and R9 are independently a saturated or partial saturated cyclic 5, 6 or 7 membered amine, imide or lactam;
or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric form.
In another preferred embodiment, the present invention is concerned with compounds wherein X is sulphur.
In another preferred embodiment, the present invention is concerned with compounds wherein R1 is COOR3 and R2 is hydrogen; wherein R3 is defined as above.
In another preferred embodiment, the present invention is concerned with compounds wherein n and m are 1.
In another preferred embodiment, the present invention is concerned with compounds wherein R5 is C1-C6alkylNR8R9.
In another preferred embodiment, the present invention is concerned with compounds wherein R4 and R6 are hydrogen.
In another preferred embodiment, the present invention is concerned with compounds wherein R1 is 5-tetrazolyl, R2 is hydrogen, and R5 is C1-C6alkylNR8R9.
In another preferred embodiment, the present invention is concerned with compounds wherein R6 is C1-C6alkylNR8R9.
In another preferred embodiment, the present invention is concerned with compounds wherein R4 and R5 are hydrogen.
In another preferred embodiment, the present invention is concerned with compounds wherein R1 is 5-tetrazolyl, R2 is hydrogen, and R6 is C1-C6alkylNR8R9.
In another preferred embodiment, the present invention is concerned with compounds wherein R5 and R6 are C1-C6alkylNR8R9.
In another preferred embodiment, the present invention is concerned with compounds wherein R1 is COOR3 and R2 is hydrogen; wherein R3 is defined as above.
In another preferred embodiment, the present invention is concerned with compounds wherein R1 is 5-tetrazolyl.
In another preferred embodiment, the present invention is concerned with compounds wherein R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with two oxo groups.
In another preferred embodiment, the present invention is concerned with compounds wherein the ring system is isoindolyl.
In another preferred embodiments, the present invention is concerned with compounds wherein R7 is C1-C6alkoyxcarbonyl.
In another preferred embodiment, the present invention is concerned with compounds wherein R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with two oxo groups.
In another preferred embodiment, the present invention is concerned with compounds wherein the ring system is isoindolyl.
In another preferred embodiment, the present invention is concerned with compounds wherein R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 7 carbon atoms and one sulfur atom, the ring system being optionally substituted with three oxo groups.
In another preferred embodiment, the present invention is concerned with compounds wherein the ring system is 2,3-dihydro-benzo[d]isothiazoly.
In another preferred embodiment, the present invention is concerned with compounds wherein R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 7 carbon atoms and one sulfur atom, the ring system being optionally substituted with two oxo groups.
In another preferred embodiment, the present invention is concerned with compounds wherein the ring system is 2,3-dihydro-benzo[d]isothiazoly.
In another preferred embodiment, the present invention is concerned with compounds wherein R7 is C1-C6alkoxycarbonyl.
In another preferred embodiment, the present invention is concerned with compounds wherein R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with one oxo group.
In another preferred embodiment, the present invention is concerned with compounds wherein the ring system is optionally substituted isoindolyl.
In another preferred embodiment, the present invention is concerned with compounds wherein the ring system is optionally substituted 1-oxo-1,3-dihydro-isoindolyl.
In another preferred embodiment, the present invention is concerned with compounds wherein R7 is C1-C6alkoxycarbonyl.
In another preferred embodiment, the present invention is concerned with compounds wherein R5 and R6 are C1-C6alkylNR8R9.
In another preferred embodiment, the present invention is concerned with compounds of Formula I wherein R5 is 1,3-dihydro-isoindol, substituted with 1 or 2 oxo groups at the atom positions adjacent to the nitrogen atom and optionally substituted with hydroxy, C1-6-alkyloxy, arylC1-6-alkyloxy or C1-6-alkylcarboxy, and wherein R7 is hydrogen, alkyl, alkyloxycarbonyl, arylalkyl or aryl wherein aryl is optionally substituted with methoxy.
In another preferred embodiment, the present invention is concerned with compounds of Formula I wherein R5 is 1,1,3-trioxo-1,2-dihydro-1H-benzo[d]isothiazol-2-yl and wherein R7 is hydrogen or arylalkyl.
In another preferred embodiment, the present invention is concerned with compounds of Formula I wherein R5 or R6 is arylaminoalkyl, wherein aryl is 1,1-dioxo-1,2-dihydro-1H-benzo[d]isothiazol-3-yl.
In another preferred embodiment, the present invention is concerned with compounds of Formula I wherein R5 or R6 is arylcarbonylaminoalkyl, wherein aryl is phenyl, indol-3-yl, indol-2-yl, 1,2,3-triazol4-yl, quinolin-4-yl or naphth-1-yl wherein aryl is optionally substituted, and wherein R7 is hydrogen or arylalkyl optionally substituted with methoxy.
In another preferred embodiment, the present invention is concerned with compounds of Formula I wherein R5 is arylalkylaminoalkyl wherein aryl is phenyl, dibenzofuranyl, naphth-2-yl or indo-3-yl, and wherein alkyl and aryl are optionally substituted, and wherein R7 is hydrogen or arylalkyl optionally substituted with methoxy.
In another preferred embodiment, the present invention is concerned with compounds of Formula I wherein R6 is alkylNR8R9, wherein R8 is alkylcarbonyl and R9 is arylalkyl, wherein aryl is optionally substituted.
In a preferred embodiment of the invention X in formula 1 is sulphur. In another preferred embodiment of the invention R1 is COOR3 and R2 is hydrogen; wherein R3 is hydrogen, C1-C6alkyl or arylC1-C6alkyl.
In a further preferred embodiment of the invention n and m are 1.
In a further preferred embodiment of the invention R4 and R6 are both hydrogen and R5 is C1-C6alkylNR8R9 and R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with two oxo groups.
Most preferred are R8 and R9 together with the nitrogen to which they are attached forming an isoindolyl-1,3-dione optionally substituted.
In another preferred embodiment of the invention R4 and R5 are both hydrogen and R6 is C1-C6alkylNR8R9 and R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with three oxo groups.
Most preferred are R8 and R9 together with the nitrogen to which they are attached forming an 1,1-dioxo-1,2-dihydro-1H-benzo[d]isothiazolyl-3-one optionally substituted.
In a further preferred embodiment of the invention R4 and R5 are both hydrogen and R6 is C1-C6alkylNR8R9 and R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with two oxo groups.
Most preferred are R8 and R9 together with the nitrogen to which they are attached forming an isoindolyl-1,3-dione optionally substituted.
In a further preferred embodiment of the invention R4 and R6 are both hydrogen and R5 is C1-C6alkylNR8R9 and R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with one oxo group.
Most preferred are R8 and R9 together with the nitrogen to which they are attached forming an optionally substituted 1-oxo-1,3-dihydro-isoindolyl ring.
In another preferred embodiment of the invention R4 and R5 are both hydrogen and R6 is C1-C6alkylNR8R9 and R8 and R9 are together with the nitrogen to which they are attached forming a partially saturated bicyclic ring system containing 8 carbon atoms, the ring system being optionally substituted with one oxo group.
Most preferred are R8 and R9 together with the nitrogen to which they are attached forming an optionally substituted 1-oxo-1,3-dihydro-isoindolyl ring.
In a preferred embodiment of the invention R7 is C1-C6alkoxycarbonyl.
The following compounds are preferred:
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3,6-dicarboxylic acid 6-ethyl ester;
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
(L)-5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Hydroxy-1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thien [2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-7-(1,1,3-trioxo-1,3-dihydro-1H-benzo[d]isothiazol-2-ylmethyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(1,1-Dioxo-1,3-dihydro-1H-benzo[d]isothiazol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
or a pharmaceutically acceptable salt thereof.
The following compounds are also preferred:
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3,6-dicarboxylic acid 6-ethyl ester;
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
(S)-5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Hydroxy-1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno [2,3-c]pyridine-3-carboxylic acid;
5-(4-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-((1,1-Dioxo-1H-benzo[d]isothiazol-3-ylamino)-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-((1,1-Dioxo-1H-benzo[d]isothiazol-3-ylamino)-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Methoxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Benzyloxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(7-Methoxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(R)-(7-Methoxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Benzyloxy-1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(6-Methoxy-4-methoxycarbonyl-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(1,1,3-trioxo-1,3-dihydro-1H-benzo[d]isothiazol-2-ylmethyl)-4,7-dihydro-5H-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-7-(1,1,3-trioxo-1,3-dihydro-1H-benzo[d]isothiazol-2-ylmethyl)-4,7-dihydro-5H-thieno[2,3-c]pyridine-3-carboxylic acid;
7-Carbamoyl-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(2-oxo-tetrahydro-thiophen-3-ylcarbamoyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-phenylcarbamoyl-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-7-phenylcarbamoyl-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
or a pharmaceutically acceptable salt thereof.
The following compounds are also preferred:
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3,6-dicarboxylic acid 6-ethyl ester;
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Hydroxy-1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(4-Hydroxy-1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(S)-(1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-((1,1-Dioxo-1H-benzo[d]isothiazol-3-ylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-((1,1-Dioxo-1H-benzo[d]isothiazol-3-ylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Methoxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Benzyloxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl1)-2-(oxalyl-amino-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(7-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(((5-Benzyloxy-1H-indole-2-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(((6-Bromo-2-p-tolyl-quinoline-4-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
6-(4-Methoxy-benzyl)-7-(((5-methyl-2-phenyl-2H-[1,2,3]triazole4-carbonyl)amino)methyl)-2(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(((1H-Indole-3-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-((4-Ethoxy-2-hydroxy-benzoylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino) -4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-((4-Benzoylamino-benzoylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(((Biphenyl4-carbonyl)-amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(((1H-Indole-2-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-((3-Biphenyl4-yl-acryloylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c)pyridine-3-carboxylic acid;
6-(4-Methoxy-benzyl)-7-(((5-methoxy-1H-indole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-((4-Benzyl-benzoylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
6-(4-Methoxy-benzyl)-7-(((naphthalene-1-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
6-(4-Methoxy-benzyl)-5-((2-naphthalen-2-yl-ethylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-((2-Benzo[1,3]dioxol-5-yl-acetylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-((2-Dibenzofuran-2-yl-ethyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
6-(4-Methoxy-benzyl)-5-((2-(5-methoxy-2-methyl-1H-indol-3-yl)-acetylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-((2-(1H-Indol-3-yl)-2-oxo-acetylamino)methyl)-2-(Oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(R)-(7-Methoxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(7-Methoxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(4-Hydroxy-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(S)-((4-phenoxy-benzylamino)methyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-((4-Acetylamino-benzylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(S)-((Acetyl-(4-phenoxy-benzyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(S)-((Acetyl-benzyl-amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-((1,1-Dioxo-1H-benzo[d]isothiazol-3-ylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(4-Benzyloxy-1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(6-Methoxy-4-methoxycarbonyl-1-oxo-1,3-dihydro-isoindol-2-ylmethyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(1,1,3-trioxo-1,3-dihydro-1H-benzo[d]isothiazol-2-ylmethyl)-4,7-dihydro-5H-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-7-(1,1,3-trioxo-1,3-dihydro-1H-benzo[d]isothiazol-2-ylmethyl)-4,7-dihydro-5H-thieno[2,3-c]pyridine-3-carboxylic acid;
7-(R)-Carbamoyl-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(S)-(2-oxo-tetrahydro-thiophen-3-ylcarbamoyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(S)-phenylcarbamoyl4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-7-(R)-phenylcarbamoyl-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(R),7-(R)-Bis-benzyloxymethyl-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
6-Benzyl-2-(oxalyl-amino)-5-(1,1,3-trioxo-1,3-dihydro-1,6-benzo[d]isothiazol-2-ylmethyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric form, or prodrug thereof.
The compounds are evaluated for biological activity with a truncated form of PTPI B (corresponding to the first 321 amino acids), which was expressed in E. coli and purified to apparent homogeneity using published procedures well-known to those skilled in the art. The enzyme reactions are carried out using standard conditions essentially as described by Burke et al. (Biochemistry 35; 15989-15996 (1996)). The assay conditions are as follows. Appropriate concentrations of the compounds of the invention are added to the reaction mixtures containing different concentrations of the substrate, p-nitrophenyl phosphate (range: 0.16 to 10 mMxe2x80x94final assay concentration). The buffer used was 50 mM HEPES pH 7.0, 100 mM sodium chloride, 0.1% (w/v) bovine serum albumin, 5 mM glutathione, and 1 mM EDTA. The reaction was started by addition of the enzyme and carried out in microtiter plates at 25xc2x0 C. for 60 minutes. The reactions are stopped by addition of NaOH. The enzyme activity was determined by measurement of the absorbance at 405 nm with appropriate corrections for absorbance at 405 nm of the compounds and p-nitrophenyl phosphate. The data are analyzed using nonlinear regression fit to classical Michaelis Menten enzyme kinetic models. Inhibition is expressed as Ki values in nM. The results of representative experiments are shown in Table 1.
Analysis for Blood Glucose Lowering Effects
The compounds of the invention are tested for blood glucose lowering effects in diabetic, obese female ob/ob mice. The mice are of similar age and body weights and they are randomized into groups of ten mice. They have free access to food and water during the experiment. The compounds are administered by either by gavage, subcutaneous, intravenous or intraperitoneal injections. The control group receives the same volume of vehicle as the mice that receive the compounds. Non-limiting examples of dose-range: 0.1, 0.3, 1.0, 3.0, 10, 30, 100 mg per kg body weight. The blood glucose levels are measured two times before administration of the compounds of the invention and vehicle (to the control group). After administration of the compound, the blood glucose levels are measured at the following time points: 1, 2, 4, 6, and 8 hours. A positive response is defined either as (i) a more than 25 percent reduction in blood glucose levels in the group receiving the compound of the invention compared to the group receiving the vehicle at any time point or (ii) statistically significant (i.e. p less than 0.05) reduction in the area under the blood glucose curve during the whole period (i.e. 8 hrs) in the group treated with the compounds of the invention compared to the group receiving the vehicle.
Compounds that show positive response can be used as development candidates and used for treatment of human diseases such as diabetes and obesity.
In accordance with one aspect of the invention, the compounds of the invention are prepared as illustrated in the following reaction scheme: 
a) NCCH2COOR3, sulphur, morpholine or triethylamine, ethanol; b) R3OCOCOimidazole, tetrahydrofuran; c) 25% trifluoroacetic acid/dichloromethane; wherein n, m, X, R1, R2, R3, R4, R5, R6 and R7 are defined above;
When R4 is hydrogen the reaction step a) in Method A gives a mixture of regioisomers which can be separated by use of column chromatography known to thus skilled in the art. 
By allowing an amine (I) and a substituted oxalylamide (II) to react under basic conditions (e.g. K2CO3, in N,N-dimethylformamide or methylethylketone) or under Mitsunobu conditions (Oyo Mitsunobu, Synthesis, (1981) 1-28) to yield (III) wherein W is OH, OSO2Me or halo, and n, m, X, R1, R2, R3, R4, R6, R7 and R8 are defined above. 
By allowing an amine (I) and a substituted oxalylamide (II) to react under basic conditions (e.g. K2CO3, in N,N-dimethylformamide or methylethylketone) or under Mitsunobu conditions (Oyo Mitsunobu, Synthesis, (1981) 1-28) to yield (III) wherein W is OH, OSO2Me or halo, and n, m, X, R1, R2, R3, R4, R5, R7 and R8 are defined above.
Pharmacological Preparations
In another aspect, the present invention includes within its scope pharmaceutical compositions comprising, as an active ingredient, at least one of the compounds of the general formula I or a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier or diluent.
The present compounds may also be administered in combination with one or more further pharmacologically active substances e.g., selected from antiobesity agents, antidiabetics, antihypertensive agents, agents for the treatment and/or prevention of complications resulting from or associated with diabetes and agents for the treatment and/or prevention of complications and disorders resulting from or associated with obesity.
Thus, in a further aspect of the invention the present compounds may be administered in combination with one or more antiobesity agents or appetite regulating agents.
Such agents may be selected from the group consisting of CART (cocaine amphetamine regulated transcript) agonists, NPY (neuropeptide Y) antagonists, MC4 (melanocortin 4) agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF (corticotropin releasing factor) agonists, CRF BP (corticotropin releasing factor binding protein) antagonists, urocortin agonists, xcex23 agonists, MSH (melanocyte-stimulating hormone) agonists, MCH (melanocyte-concentrating hormone) antagonists, CCK (cholecystokinin) agonists, serotonin re-uptake inhibitors, serotonin and noradrenaline re-uptake inhibitors, mixed serotonin and noradrenergic compounds, 5HT (serotonin) agonists, bombesin agonists, galanin antagonists, growth hormone, growth hormone releasing compounds, TRH (thyreotropin releasing hormone) agonists, UCP 2 or 3 (uncoupling protein 2 or 3) modulators, leptin agonists, DA agonists (bromocriptin, doprexin), lipase/amylase inhibitors, PPAR (peroxisome proliferator activated receptor) modulators, RXR (retinoid X receptor) modulators or TR xcex2 agonists.
In one embodiment of the invention the antiobesity agent is leptin.
In another embodiment the antiobesity agent is dexamphetamine or amphetamine.
In another embodiment the antiobesity agent is fenfluramine or dexfenfluramine.
In still another embodiment the antiobesity agent is sibutramine.
In a further embodiment the antiobesity agent is orlistat.
In another embodiment the antiobesity agent is mazindol or phentermine.
Suitable antidiabetics comprise insulin, GLP-1 (glucagon like peptide-1) derivatives such as those disclosed in WO 98/08871 to Novo Nordisk A/S, which is incorporated herein by reference as well as orally active hypoglycaemic agents.
The orally active hypoglycaemic agents preferably comprise sulphonylureas, biguanides, meglitinides, oxadiazolidinediones, thiazolidinediones, glucosidase inhibitors, glucagon antagonists such as those disclosed in WO 99/01423 to Novo Nordisk A/S and Agouron Pharmaceuticals, Inc., GLP-1 agonists, potassium channel openers such as those disclosed in WO 97/26265 and WO 99/03861 to Novo Nordisk A/S which are incorporated herein by reference, insulin sesitizers, DPP-IV (dipeptidyl peptidase-IV) inhibitors, inhibitors of hepatic enzymes involved in stimulation of gluconeogenesis and/or glycogenolysis, glucose uptake modulators, compounds modifying the lipid metabolism such as antihyperlipidemic agents and antilipidemic agents as HMG CoA inhibitors (statins), compounds lowering food intake, PPAR and RXR agonists and agents acting on the ATP-dependent potassium channel of the xcex2-cells.
In one embodiment of the invention the present compounds are administered in combination with insulin.
In a further embodiment the present compounds are administered in combination with a sulphonylurea e.g. tolbutamide, glibenclamide, glipizide or glicazide.
In another embodiment the present compounds are administered in combination with a biguanide e.g. metformin.
In yet another embodiment the present compounds are administered in combination with a meglitinide e.g. repaglinide.
In still another embodiment the present compounds are administered in combination with a thiazolidinedione e.g. troglitazone, ciglitazone, pioglitazone, rosiglitazone or compounds disclosed in WO 97/41097 such as 5-[[4-[3-Methyl-4-oxo-3,4-dihydro-2-quinazolinyl]methoxy]phenyl-methyl]thiazolidine-2,4-dione or a pharmaceutically acceptable salt thereof, preferably the potassium salt.
Furthermore, the present compounds may be administered in combination with the insulin sensitizers disclosed in WO 99/19313 such as (-) 3-[4-[2-Phenoxazin-10-yl)ethoxy]phenyl]-2-ethoxypropanoic acid or a pharmaceutically acceptable salts thereof, preferably the arginine salt.
In a further embodiment the present compounds are administered in combination with an xcex1-glucosidase inhibitor e.g. miglitol or acarbose.
In another embodiment the present compounds are administered in combination with an agent acting on the ATP-dependent potassium channel of the xcex2-cells e.g. tolbutamide, glibenclamide, glipizide, glicazide or repaglinide.
Furthermore, the present compounds may be administered in combination with nateglinide.
In still another embodiment the present compounds are administered in combination with an antihyperlipidemic agent or antilipidemic agent e.g. cholestyramine, colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextrothyroxine.
In a further embodiment the present compounds are administered in combination with more than one of the above-mentioned compounds e.g. in combination with a sulphonylurea and metformin, a sulphonylurea and acarbose, repaglinide and metformin, insulin and a sulphonylurea, insulin and metformin, insulin, insulin and lovastatin, etc.
Furthermore, the present compounds may be administered in combination with one or more antihypertensive agents. Examples of antihypertensive agents are xcex2-blockers such as alprenolol, atenolol, timolol, pindolol, propranolol and metoprolol, ACE (angiotensin converting enzyme) inhibitors such as benazepril, captopril, enalapril, fosinopril, lisinopril, quinapril and ramipril, calcium channel blockers such as nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and xcex1-blockers such as doxazosin, urapidil, prazosin and terazosin. Further reference can be made to Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, Pa., 1995.
It should be understood that any suitable combination of the compounds according to the invention with one or more of the above-mentioned compounds and optionally one or more further pharmacologically active substances are considered to be within the scope of the present invention.
For the above indications the dosage will vary depending on the compound of the invention employed, on the mode of administration and on the therapy desired. However, in general, satisfactory results are obtained with a dosage of from about 0.5 mg to about 1000 mg, preferably from about 1 mg to about 500 mg of compounds of the invention, conveniently given from 1 to 5 times daily, optionally in sustained release form. Usually, dosage forms suitable for oral administration comprise from about 0.5 mg to about 1000 mg, preferably from about 1 mg to about 500 mg of the compounds of the invention admixed with a pharmaceutical carrier or diluent.
The compounds of the invention may be administered in a pharmaceutically acceptable acid addition salt form or where possible as a metal or a C1-6-alkylammonium salt. Such salt forms exhibit approximately the same order of activity as the free acid forms.
This invention also relates to pharmaceutical compositions comprising a compound of the invention or a pharmaceutically acceptable salt thereof and, usually, such compositions also contain a pharmaceutical carrier or diluent. The compositions containing the compounds of this invention may be prepared by conventional techniques and appear in conventional forms, for example capsules, tablets, solutions or suspensions.
The pharmaceutical carrier employed may be a conventional solid or liquid carrier. Examples of solid carriers are lactose, terra alba, sucrose, talc, gelatine, agar, pectin, acacia, magnesium stearate and stearic acid. Examples of liquid carriers are syrup, peanut oil, olive oil and water.
Similarly, the carrier or diluent may include any time delay material known to the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.
If a solid carrier for oral administration is used, the preparation can be tabletted, placed in a hard gelatine capsule in powder or pellet form or it can be in the form of a troche or lozenge. The amount of solid carrier will vary widely but will usually be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.
Generally, the compounds of this invention are dispensed in unit dosage form comprising 10-200 mg of active ingredient in or together with a pharmaceutically acceptable carrier per unit dosage.
The dosage of the compounds according to this invention is 1-500 mg/day, e.g. about 100 mg per dose, when administered to patients, e.g. humans, as a drug.
A typical tablet that may be prepared by conventional tabletting techniques contains
* Acylated monoglyceride used as plasticiser for film coating. 
The route of administration may be any route, which effectively transports the active compound to the appropriate or desired site of action, such as oral or parenteral e.g. rectal, transdermal, subcutaneous, intranasal, intramuscular, topical, intravenous, intraurethral, ophthalmic solution or an ointment, the oral route being preferred.